摘要
Objective To observe the effect of the transfer of multidrug resistance gene (mdr1) into human hematopoietic progenitor cells (HPC) on the chemoprotection Methods Human CD34 + cells served as a target of mdr1 gene transfer Retroviral vector SF mdr containing human total length mdr1cDNA was introduced into packing cells GP envAM12 by liposome mediated transfection The mdr1 gene was transduced into human CD34 + cells by retroviral supernatants of packing cells The integration and expression of the mdr1 gene and its protein (P170) in transduced cells were determined by PCR, RT PCR, and flow cytometry The drug resistance of chemotherapy in transduced HPC was determined by culturing colonies Results The mdr1 gene was integrated and expressed in transduced CD34 + cells The efficiency of mdr1 gene transfer was 10%-14% Compared with untransduced controls, within a certain range of drug concentration, the number of drug resistant colony in transduced HPC for taxol, doxorubicin,VCR and VP16 were increased by 3 6±2 1 fold, 2 9±0 3 fold, 1 9±0 4 fold, and 3 5±0 5 fold, respectively Conclusion The transfer of the mdr1 gene into human HPC can increase the drug resistance of the transduced cells to corresponding chemotherapeutic drugs that may provide some degree of chemoprotection for HPC
Objective To observe the effect of the transfer of multidrug resistance gene (mdr1) into human hematopoietic progenitor cells (HPC) on the chemoprotection Methods Human CD34 + cells served as a target of mdr1 gene transfer Retroviral vector SF mdr containing human total length mdr1cDNA was introduced into packing cells GP envAM12 by liposome mediated transfection The mdr1 gene was transduced into human CD34 + cells by retroviral supernatants of packing cells The integration and expression of the mdr1 gene and its protein (P170) in transduced cells were determined by PCR, RT PCR, and flow cytometry The drug resistance of chemotherapy in transduced HPC was determined by culturing colonies Results The mdr1 gene was integrated and expressed in transduced CD34 + cells The efficiency of mdr1 gene transfer was 10%-14% Compared with untransduced controls, within a certain range of drug concentration, the number of drug resistant colony in transduced HPC for taxol, doxorubicin,VCR and VP16 were increased by 3 6±2 1 fold, 2 9±0 3 fold, 1 9±0 4 fold, and 3 5±0 5 fold, respectively Conclusion The transfer of the mdr1 gene into human HPC can increase the drug resistance of the transduced cells to corresponding chemotherapeutic drugs that may provide some degree of chemoprotection for HPC
基金
ThisstudywassupportedbygrantsfromtheMinistryofPublicHealth,China!(No95150)